C4785

Collagenase aus Clostridium histolyticum

0.2 μm filtered, release of physiologically active rat pancreatic islets tested, Type XI-S, 2-5 FALGPA units/mg solid, >1200 CDU/mg solid

• Synonym(e): Clostridiopeptidase A
• CAS-Nummer: 9001-12-1
• UNSPSC Code: 12352204
• EC Number: 232-582-9
• MDL number: MFCD00130830
• EG-Nummer: 3.4.24.3 (BRENDA, IUBMB)

Eigenschaften

• Quality Segment: 200
• sterility: 0.2 μm filtered
• form: lyophilized powder
• specific activity: >1200 CDU/mg solid, 2-5 FALGPA units/mg solid
• mol wt: 68-130 kDa
• suitability: release of physiologically active rat pancreatic islets tested
• storage temp.: −20°C

Beschreibung

Biochem/physiol Actions

Collagenase wird durch vier Grammatom Calcium pro Mol Enzym aktiviert. Sie wird durch Ethylenglycol-bis-(beta-aminoethylether)-N, N, N′,N′-tetraessigsäure, β-Mercaptoethanol, Glutathion, Thioglycolsäure und 8-Hydroxychinolin gehemmt.

Effective release of cells from tissue requires the action of collagenase enzymes and the neutral protease. Collagenase is activated by four gram atom calcium (Ca²⁺) per mole enzyme. The culture filtrate is thought to contain at least 7 different proteases ranging in molecular weight from 68-130 kDa. The pH optimum is 6.3-8.8. The enzyme is typically used to digest the connective components in tissue samples to liberate individual cells. Collagenase treatment can cause some cells to die. Typically, concentrations varying from 0.1 to 5 mg/mL are used for digestion. The duration of reaction varies from 15 minutes to several hours and yields a satisfactory efficiency of cell dissociation without causing too much cell death. Krebs Ringer buffer with calcium and BSA is preferred and Zn²⁺ is required for activity. This enzyme is tested for suitability for the release of hepatocytes (at approx. 1 mg/mL in a total volume of 100 mL) for each rat liver.

Preparation Note

Also contains clostripain, nonspecific neutral protease, and tryptic activities.

Prepared from Type XI (C7657)

Other Notes

Eine Collagen-Aufschlusseinheit (Collagen Digestion Unit, CDU) setzt Peptide aus Rinder-Achillessehnencollagen entsprechend der Ninhydrin-Anfärbung von 1,0 μmol Leucin in 5 Stunden bei pH 7,4 und 37 °C in Gegenwart von Calciumionen frei. Eine FALGPA-Hydrolyseeinheit hydrolysiert 1,0 μmol Furylacryloyl-Leu-Gly-Pro-Ala pro Minute bei 25 °C. Eine neutrale Protease-Einheit hydrolysiert die Caseinmenge, die eine Färbung entsprechend 1,0 μmol Tyrosin pro 5 Stunden bei pH 7,5 und 37 °C produziert. Eine Clostripain-Einheit hydrolysiert 1,0 μmol BAEE pro Minute bei pH 7,6 und 25 °C in Gegenwart von DTT.

Sicherheitshinweise

• pictograms: GHS08,GHS07
• signalword: Danger
• hcodes: H315,H319,H334,H335
• pcodes: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• target_organs: Respiratory system
• Lagerklasse: 11 - Combustible Solids
• wgk: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable
• ppe: dust mask type N95 (US), Eyeshields, Faceshields, Gloves